RESUMEN
Self-assembling conjugated polymer nanoparticles containing PVK and PLGA-PEG as a matrix polymer were doped with both a luminescent rare-earth complex and magnetic nanoparticles (SPIONs), giving rise to materials that are both luminescent and magnetic. Nanoparticle sizes ranged from 80-110 nm without SPIONs and showed an increase in size (200-1000 nm) with additional SPION content (11-54%). Quantum yields (QYs) of 24% and 18% were measured for systems without and with 11% SPIONs, respectively. Optical properties were stable and suitable for biological imaging applications.
Asunto(s)
Europio/química , Nanopartículas de Magnetita/química , Imagen Óptica , Poliésteres/química , Polietilenglicoles/química , Investigación Biomédica , Células Cultivadas , Humanos , Hidrodinámica , Microscopía Confocal , Tamaño de la Partícula , Teoría Cuántica , Propiedades de Superficie , Linfocitos T/citologíaRESUMEN
Acute and chronic respiratory diseases account for major causes of illness and deaths worldwide. Recent developments of biotherapeutics opened a new era in the treatment and management of patients with respiratory diseases. When considering the delivery of therapeutics, the inhaled route offers great promises with a direct, non-invasive access to the diseased organ and has already proven efficient for several molecules. To assist in the future development of inhaled biotherapeutics, experimental models are crucial to assess lung deposition, pharmacokinetics, pharmacodynamics and safety. This review describes the animal models used in pulmonary research for aerosol drug delivery, highlighting their advantages and limitations for inhaled biologics. Overall, non-clinical species must be selected with relevant scientific arguments while taking into account their complexities and interspecies differences, to help in the development of inhaled medicines and ensure their successful transposition in the clinics.
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Aerosoles/administración & dosificación , Preparaciones Farmacéuticas/administración & dosificación , Terapia Respiratoria/métodos , Administración por Inhalación , Animales , Sistemas de Liberación de Medicamentos/métodos , Humanos , Modelos AnimalesRESUMEN
This study compares the performance of a microfluidic technique and a conventional bulk method to manufacture conjugated polymer nanoparticles (CPNs) embedded within a biodegradable poly(ethylene glycol) methyl ether-block-poly(lactide-co-glycolide) (PEG5K-PLGA55K) matrix. The influence of PEG5K-PLGA55K and conjugated polymers cyano-substituted poly(p-phenylene vinylene) (CN-PPV) and poly(9,9-dioctylfluorene-2,1,3-benzothiadiazole) (F8BT) on the physicochemical properties of the CPNs was also evaluated. Both techniques enabled CPN production with high end product yields (â¼70-95%). However, while the bulk technique (solvent displacement) under optimal conditions generated small nanoparticles (â¼70-100 nm) with similar optical properties (quantum yields â¼35%), the microfluidic approach produced larger CPNs (140-260 nm) with significantly superior quantum yields (49-55%) and tailored emission spectra. CPNs containing CN-PPV showed smaller size distributions and tuneable emission spectra compared to F8BT systems prepared under the same conditions. The presence of PEG5K-PLGA55K did not affect the size or optical properties of the CPNs and provided a neutral net electric charge as is often required for biomedical applications. The microfluidics flow-based device was successfully used for the continuous preparation of CPNs over a 24 hour period. On the basis of the results presented here, it can be concluded that the microfluidic device used in this study can be used to optimize the production of bright CPNs with tailored properties with good reproducibility.
RESUMEN
Red emitting materials are desirable in biology due to the transparency of certain biological tissues at these wavelengths. Here, we report the synthesis of aqueous dispersions of amphiphilic protein (hydrophobin) capped red-emitting cyano-substituted poly(p-phenylenevinylene) conjugated polymer nanoparticles (CPNs) and their use in labeling live mammalian (HeLa) cells.
RESUMEN
The development of simple routes to emissive solid-state materials is of paramount interest, and in this report we describe the biosynthesis of infrared emitting quantum dots in a living plant via a mutual antagonistic reaction. Exposure of common Allium fistulosum to mercury and tellurium salts under ambient conditions resulted in the expulsion of crystalline, non-passivated HgTe quantum dots that exhibited emissive characteristics in the near-infrared spectral region, a wavelength range that is important in telecommunications and solar energy conversion.
Asunto(s)
Allium/química , Rayos Infrarrojos , Puntos Cuánticos/químicaRESUMEN
The development of clinically acceptable albumin-based nanoparticle formulations for use in pulmonary drug delivery has been hindered by concerns about the toxicity of nanomaterials in the lungs combined with a lack of information on albumin nanoparticle clearance kinetics and biodistribution. In this study, the in vivo biocompatibility of albumin nanoparticles was investigated following a single administration of 2, 20, and 390µg/mouse, showing no inflammatory response (TNF-α and IL-6, cellular infiltration and protein concentration) compared to vehicle controls at the two lower doses, but elevated mononucleocytes and a mild inflammatory effect at the highest dose tested. The biodistribution and clearance of (111)In labelled albumin solution and nanoparticles over 48h following a single pulmonary administration to mice was investigated by single photon emission computed tomography and X-ray computed tomography imaging and terminal biodistribution studies. (111)In labelled albumin nanoparticles were cleared more slowly from the mouse lung than (111)In albumin solution (64.1±8.5% vs 40.6±3.3% at t=48h, respectively), with significantly higher (P<0.001) levels of albumin nanoparticle-associated radioactivity located within the lung tissue (23.3±4.7%) compared to the lung fluid (16.1±4.4%). Low amounts of (111)In activity were detected in the liver, kidneys, and intestine at time points >24h indicating that small amounts of activity were cleared from the lungs both by translocation across the lung mucosal barrier, as well as mucociliary clearance. This study provides important information on the fate of albumin vehicles in the lungs, which may be used to direct future formulation design of inhaled nanomedicines.
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Sistemas de Liberación de Medicamentos , Nanopartículas , Albúmina Sérica Bovina/farmacocinética , Administración por Inhalación , Animales , Líquido del Lavado Bronquioalveolar/citología , Recuento de Células , Interleucina-6/metabolismo , Mucosa Intestinal/metabolismo , Riñón/metabolismo , Hígado/embriología , Pulmón/metabolismo , Masculino , Ratones Endogámicos BALB C , Nanopartículas/administración & dosificación , Óxido Nítrico/metabolismo , Albúmina Sérica Bovina/administración & dosificación , Albúmina Sérica Bovina/farmacología , Distribución Tisular , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
The synthesis of designer solid-state materials by living organisms is an emerging field in bio-nanotechnology. Key examples include the use of engineered viruses as templates for cobalt oxide (Co(3)O(4)) particles, superparamagnetic cobalt-platinum alloy nanowires and gold-cobalt oxide nanowires for photovoltaic and battery-related applications. Here, we show that the earthworm's metal detoxification pathway can be exploited to produce luminescent, water-soluble semiconductor cadmium telluride (CdTe) quantum dots that emit in the green region of the visible spectrum when excited in the ultraviolet region. Standard wild-type Lumbricus rubellus earthworms were exposed to soil spiked with CdCl(2) and Na(2)TeO(3) salts for 11 days. Luminescent quantum dots were isolated from chloragogenous tissues surrounding the gut of the worm, and were successfully used in live-cell imaging. The addition of polyethylene glycol on the surface of the quantum dots allowed for non-targeted, fluid-phase uptake by macrophage cells.
Asunto(s)
Biotecnología/métodos , Sustancias Luminiscentes/metabolismo , Nanotecnología/métodos , Oligoquetos/metabolismo , Puntos Cuánticos , Animales , Compuestos de Cadmio/metabolismo , Línea Celular , Exposición a Riesgos Ambientales , Tracto Gastrointestinal/química , Tracto Gastrointestinal/citología , Histocitoquímica , Sustancias Luminiscentes/química , Sustancias Luminiscentes/aislamiento & purificación , Macrófagos/metabolismo , Ratones , Oligoquetos/química , Tamaño de la Partícula , Telurio/metabolismoRESUMEN
Cyclodextrins (CDs) are promising solubility enhancers for inhaled drug delivery. However, they have dose-dependent effects on the respiratory epithelium, which may have advantages for permeability enhancement but also gives rise to safety concerns. In this study, the methyl thiazol tetrazolium (MTT) assay was used to compare a new sparingly methylated beta-CD, Kleptose Crysmebeta (Crysmeb) with the more established CD derivatives hydroxypropyl-gamma-cyclodextrin (HPgammaCD), randomly methylated beta-cyclodextrin (Rameb) and hydroxypropyl-beta-cyclodextrin (HPbetaCD). The betaCD derivatives affected cell metabolism in A549 cells in a concentration dependent manner with LD(50) of 56, 31 and 11 mM obtained for HPbetaCD, Crysmeb and Rameb, respectively. Calu-3 cells were less susceptible to betaCD with an LD(50) of 25 mM being obtained for Rameb only. Permeability increases in Calu-3 cell layers were observed with betaCD derivatives and a concentration dependency shown. The mechanism of permeability enhancement and its reversibility was investigated. Rameb produced an irreversible loss of cell layer barrier function at > or = 25 mM, but perturbations of epithelial integrity were moderate and reversible in the case of HPbetaCD and Crysmeb (25-50 mM). Given its high solubilisation capacity, the low toxicity and transient absorption promoting properties, this study identifies Crysmeb as a promising adjuvant in formulations for inhalation.
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Citotoxinas/toxicidad , Células Epiteliales/metabolismo , Mucosa Respiratoria/metabolismo , beta-Ciclodextrinas/toxicidad , Línea Celular , Células Epiteliales/efectos de los fármacos , Células Epiteliales/patología , Humanos , Metilación/efectos de los fármacos , Permeabilidad/efectos de los fármacos , Mucosa Respiratoria/efectos de los fármacos , Mucosa Respiratoria/patologíaRESUMEN
Particulate nanocarriers have been praised for their advantageous drug delivery properties in the lung, such as avoidance of macrophage clearance mechanisms and long residence times. However, instilled non-biodegradable polystyrene nanospheres with small diameters and thus large surface areas have been shown to induce pulmonary inflammation. This study examines the potential of biodegradable polymeric nanoparticles composed of poly(lactic-co-glycolic acid) (PLGA) and the novel PLGA derivative, diethylaminopropylamine polyvinyl alcohol-grafted-poly(lactic-co-glycolic acid) (DEAPA-PVAL-g-PLGA), to provoke inflammatory responses in the murine lung after intratracheal instillation. Lactate dehydrogenase (LDH) release, protein concentration, MIP-2 mRNA induction, and polymorphonucleocyte (PMN) recruitment in the bronchial alveolar lavage fluid (BALF) were used to evaluate an inflammatory response in Balb-C mice. Two sizes of polystyrene (PS) nanospheres (diameters: 75 nm and 220 nm) were included in the study for comparison. All nanoparticle suspensions were instilled at concentrations of 1 microg/microl and 2.5 microg/microl, representative of an estimated "therapeutic dose" and a concentrated "dose" of particles. In all experiments, the 75 nm PS particles exhibited elevated responses for the inflammatory markers investigated. In contrast, biodegradable particles of comparable hydrodynamic diameter showed a significantly lower inflammatory response. The most marked differences were observed in the extent of PMN recruitment. While the 75 nm and 220 nm PS nanospheres exhibited 41 and 74% PMN within the total BALF cell population after 24 h, respectively, PMN recruiting in lungs instilled with both types of biodegradable particles did not exceed values of the negative isotonic glucose control. In conclusion, evidence suggests that biodegradable polymeric nanoparticles designed for pulmonary drug delivery may not induce the same inflammatory response as non-biodegradable polystyrene particles of comparable size.
Asunto(s)
Sistemas de Liberación de Medicamentos , Pulmón/efectos de los fármacos , Neumonía/etiología , Animales , Biodegradación Ambiental , Ratones , Nanotecnología , Tamaño de la PartículaRESUMEN
This study examined the effect of nebulization on the cellular uptake and transfection efficiency of polyplexes from four polyethylenimine (PEI) modifications: branched 25 kDa PEI (bPEI), linear 22 kDa PEI (linPEI), pegylated PEI (pegPEI) and biodegradable PEI (bioPEI). Polyplexes were aerosolized with air-jet and ultrasonic nebulizers. The aerosol was collected and used to determine complex size and zeta potential. Fluorescence-assisted cell sorting (FACS) was used to quantify the cellular association of polyplexes in primary alveolar cells (AEC), A549 cells and primary bronchial cells (BEC). Confocal laser scanning microscopic images provided information about the internalization of polyplexes. Transfection efficiencies of polyplexes were quantified via measurement of luciferase expression. All polymers were stable during nebulization, although size increases were observed after air-jet nebulization. FACS studies showed a two- to three-fold increase in polyplex association with BEC compared to A549 cells, while polyplex association with AEC was negligible. BPEI, linPEI and bioPEI polyplexes were internalized, while pegPEI polyplexes remained predominately attached to the cellular membrane. Luciferase expression was detected only in BEC and A549 cells with transfection efficiencies approximately one order of magnitude higher in BEC. All PEI modifications investigated were suitable for aerosol therapy, although cell type and polymer structure significantly influenced the uptake and transfection efficiency of the polyplexes.
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Sistemas de Liberación de Medicamentos , Técnicas de Transferencia de Gen , Polietileneimina/química , Aerosoles , Línea Celular , ADN/química , Estabilidad de Medicamentos , Células Epiteliales , Humanos , Flujometría por Láser-Doppler , Nebulizadores y Vaporizadores , Fotones , Análisis Espectral , TransfecciónRESUMEN
Polyelectrolyte complexes between DNA and polyethylenimine (PEI) are promising non-viral delivery systems for pulmonary inhalation gene therapy and thus require sufficient stability during nebulization. The structure and stability of four different PEI-DNA polyplexes, namely branched (bPEI), linear (linPEI), poly(ethylene glycol)-grafted PEI (PEGPEI), biodegradable (bioPEI) PEI with DNA, were investigated. Using atomic force microscopy, the morphology of DNA and polyplexes before and after both air-jet and ultrasonic nebulization was characterized. The influence of nebulization on physico-chemical properties, particle size and zeta potential, was studied. Efficient DNA condensation to spherical particles was achieved with bPEI (90 nm) and PEGPEI (110 nm). By contrast, incomplete DNA condensations, seen as flower structures, were observed with linPEI (110 nm) and bioPEI (105 nm). Air-jet nebulization altered the polyplex structure to a greater extent than ultrasonic nebulization and resulted mainly in smaller and non-spherical particles (30-200 nm). Ultrasonic nebulization did not change the spherical structure or particle size of the polyplexes. In particular, the shape and size of the PEGPEI polyplexes did not change. We conclude that ultrasonic nebulization is a milder aerosolization method for gene delivery systems based on PEI. Additionally, PEGPEI-DNA polyplexes seem to be more stable than their counterparts, which may be advantageous in pulmonary inhalation gene therapy.
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Sistemas de Liberación de Medicamentos , Técnicas de Transferencia de Gen , Terapia Genética/métodos , Polietileneimina/química , Aerosoles , ADN/genética , Desoxirribonucleasas/química , Electroforesis en Gel de Agar , Flujometría por Láser-Doppler , Luz , Microscopía de Fuerza Atómica , Nebulizadores y Vaporizadores , Tamaño de la Partícula , Alveolos Pulmonares/citología , Alveolos Pulmonares/efectos de los fármacos , Dispersión de Radiación , UltrasonidoRESUMEN
Different classification systems for therapeutic agents exist. The most commonly used one is the ATC Code (ATC: Anatomy, Therapeutic properties, Chemical, pharmacological properties). Here, an alternative classification system (TCAT: Target-Chemistry-Anatomy-Therapy) is proposed which refers to the molecular mechanism of action or rather, target. The main subgroups of targets are: enzymes; substrates, metabolies, proteins; receptors; ion channels; transporter molecules and systems; nucleic acids, ribosomes; physicochemical mechanisms; antigen-antibody reactions; unknown targets. This target-oriented approach may be particularly useful in teaching advanced medicinal chemistry.
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Preparaciones Farmacéuticas/clasificación , Farmacología , Animales , Humanos , Farmacología/educaciónRESUMEN
PURPOSE: This study describes the development of surfactant-free, biodegradable nanoparticle systems with varying physicochemical properties and their suitability for pulmonary application via nebulization. METHODS: Nanoparticle suspensions were formulated from the branched polyester, diethylaminopropyl amine-poly(vinyl alcohol)-grafted-poly(lactide-co-glycolide) (DEAPA-PVAL-g-PLGA) alone, as well as with increasing amounts of carboxymethyl cellulose (CMC). Particle size, zeta potential, turbidity, and morphology (atomic force microscopy) were characterized. Three formulations were chosen for further study: Cationic nanoparticles without CMC, cationic nanoparticles with CMC, and anionic nanoparticles with an excess of CMC. Nanoparticle degradation was characterized, as well as stability during nebulization. Nanoparticle-cell interactions were investigated and quantified using confocal laser scanning microscopy and fluorescence spectrometry. RESULTS: Nanoparticles ranged in size from 70-250 nm and displayed zeta potentials of +58.9 to -46.6 mV. Anionic nanoparticles showed the highest stability during nebulization. The degradation rate of each nanoparticle formulation decreased with increasing amounts of CMC. Cell association was highest among cationic nanoparticles (57% and 30%, respectively), although these were not internalized. Despite a lower rate of cell association (3%), anionic nanoparticles were internalized by A549 cells. CONCLUSIONS: Surfactant-free nanoparticles from DEAPA-PVAL-g-PLGA are versatile drug delivery systems; however, only the anionic formulations investigated were proven suitable for aerosol therapy.
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Sistemas de Liberación de Medicamentos , Ácido Láctico/química , Poliésteres/química , Poliglactina 910/química , Ácido Poliglicólico/química , Polímeros/química , Alcohol Polivinílico/química , Aerosoles , Biotransformación , Carboximetilcelulosa de Sodio , Química Farmacéutica , Estabilidad de Medicamentos , Microscopía de Fuerza Atómica , Microscopía Confocal , Microesferas , Nefelometría y Turbidimetría , Tamaño de la Partícula , Excipientes Farmacéuticos , Poliglactina 910/análogos & derivados , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Espectrometría de Fluorescencia , SuspensionesRESUMEN
Epidemiological studies demonstrate an association between increased human morbidity and mortality with exposure to air pollution particulate matter. We hypothesized that such effects may be associated with the ability of the particles to mediate generation of reactive oxygen species (ROS), either directly, via interaction with ambient oxygen or indirectly through initiation of an oxidative burst in phagocytes. To test this hypothesis, we determined 8-oxo-dG formation as a measure of direct generation of ROS, in response to particulate exposures to 2'-deoxyguanosine (dG), free and in calf thymus DNA in aerated solutions as the target molecule and cell culture, to assess the relationship between induction of oxidative damage, particulate metal content and metal bioreactivity. The HPLC-ECD technique was employed for separation and quantification of 8-oxo-dG, the most widely recognized marker of DNA oxidation. Particles used in this study include: Arizona desert dust (AZDD), coal fly ash (CFA and ECFA), oil fly ash (OFA and ROFA), and ambient air [SRM 1649 and Dusseldorf (DUSS), Germany]. The major difference between these particles is the concentration of water-soluble metals. The fly ash particulates OFA and ROFA showed a significant dose-dependent increase in dG hydroxylation to 8-oxo-dG formation over the control dG (p < 0.05), with yields 0.03 and 1.25% at the highest particulate concentration (1 mg/mL). Metal ion chelators and DMSO, a hydroxyl radical scavenger, inhibited this hydroxylation. In contrast, desert dust, coal fly ash and urban air particles induced 8-oxo-dG with yields ranging from 0.003 to 0.006%, respectively, with levels unaffected by pretreatment of the particles with metal ion chelators or addition of DMSO to the incubation mixture. When calf thymus DNA was used as a substrate, all the particles induced 8-oxo-dG in a pattern similar to that observed for dG hydroxylation, but with relatively less yield. Treatment of the particles with metal ion chelator before reacting with DNA or addition of catalase in the incubation mixture, suppressed 8-oxo-dG formation significantly (p < 0.05) in oil-derived fly ash particles only. To determine whether the oxidative responses of these particulates as shown in cell-free systems were consistent with responses using a more biologically relevant environment, human airway epithelial cells were treated with the particulates and induction of 8-oxo-dG was determined. All particles induced 8-oxo-dG in the DNA of cells above culture control, except CFA. Cells exposed to 10-400 mg/mL of ROFA for 2 h induced a dose-dependent increase in 8-oxo-dG formation. Treatment of ROFA with metal ion chelator attenuated these effects. Overall, damage enhancement by particulates in dG, calf thymus, and cellular DNA as determined by 8-oxo-dG formation under aerobic conditions is consistent with the concentration of water-soluble, not the total metal content of the particle.
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Contaminantes Atmosféricos/toxicidad , Contaminación del Aire/efectos adversos , Daño del ADN , Desoxiguanosina/análogos & derivados , Metales/toxicidad , 8-Hidroxi-2'-Desoxicoguanosina , Aire/análisis , Animales , Carbono/análisis , Carbono/toxicidad , Bovinos , Sistema Libre de Células , Quelantes/farmacología , Carbón Mineral/análisis , Carbón Mineral/toxicidad , Ceniza del Carbón , ADN/análisis , ADN/efectos de los fármacos , Desoxiguanosina/química , Desoxiguanosina/metabolismo , Desoxiguanosina/farmacología , Polvo/análisis , Humanos , Metales/análisis , Oxidación-Reducción , Material Particulado , Especies Reactivas de Oxígeno/metabolismo , Mucosa Respiratoria/efectos de los fármacosRESUMEN
Ambient particulate matter (PM) concentrations have been associated with mortality and morbidity. Diesel exhaust particles (DEP) are present in ambient urban air PM. Coexisting with DEP (and PM) is ozone (O(3)), which has the potential to react with some components of DEP. Some reports have shown increased lung injury in rats coexposed to PM and O(3), but it is unclear whether this increased injury was due to direct interaction between the pollutants or via other mechanisms. To examine whether O(3) can directly react with and affect PM bioactivity, we exposed DEP to O(3) in a cell-free in vitro system and then examined the bioactivity of the resultant DEP in a rat model of lung injury. Standard Reference Material 2975 (diesel exhaust PM) was initially exposed to 0.1 ppm O(3) for 48 h and then instilled intratracheally in Sprague-Dawley rats. Rat lung inflammation and injury was examined 24 h after instillation by lung lavage. The DEP exposed to 0.1 ppm O(3) was more potent in increasing neutrophilia, lavage total protein, and LDH activity compared to unexposed DEP. The increased DEP activity induced by the O(3) exposure was not attributable to alteration by air that was also present during the O(3) exposure. Exposure of DEP to a higher O(3) concentration (1.0 ppm) led to a decreased bioactivity of the particles. In contrast, carbon black particles, low in organic content relative to DEP, did not exhibit an increase in any of the bioactivities examined after exposure to 0.1 ppm O(3). DEP incorporated O(3) (labeled with (18)O) in a linear fashion. These data suggest that ambient concentrations of O(3) can increase the biological potency of DEP. The ozonized DEP may play a role in the induction of lung responses by ambient PM.
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Contaminantes Atmosféricos/toxicidad , Oxidantes Fotoquímicos/toxicidad , Ozono/toxicidad , Neumonía/inducido químicamente , Emisiones de Vehículos/toxicidad , Adolescente , Adulto , Contaminantes Atmosféricos/química , Animales , Bronquios/citología , Bronquios/efectos de los fármacos , Bronquios/metabolismo , Células Cultivadas , Dinoprostona/biosíntesis , Dinoprostona/inmunología , Sinergismo Farmacológico , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Humanos , Pulmón/efectos de los fármacos , Masculino , Oxidantes Fotoquímicos/química , Oxidación-Reducción , Ozono/química , Tamaño de la Partícula , Neumonía/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
Human airway epithelial cells can increase expression of both lactoferrin and ferritin after exposure to catalytically active metal. These proteins transport and store metal, with coordination sites fully complexed, and therefore can diminish the oxidative stress. The intracellular transport of lactoferrin results in a transfer of complexed metal to ferritin, where it is stored in a less reactive form. This effort to control the injurious properties of metals would be facilitated by lactoferrin receptors (LfRs) on airway epithelial cells. We tested the hypotheses that 1) LfRs exist on respiratory epithelial cells and 2) exposure to both an air pollution particle, which has abundant concentrations of metals, and individual metal salts increase the expression of LfRs. Before exposure to either the particle or metals, incubation of BEAS-2B cells with varying concentrations of 125I-labeled lactoferrin demonstrated lactoferrin binding that was saturable. Measurement of 125I-lactoferrin binding after the inclusion of 100 micrograms/ml of oil fly ash in the incubation medium demonstrated increased binding within 5 min of exposure, which reached a maximal value at 45 min. Inclusion of 1.0 mM deferoxamine in the incubation of BEAS-2B cells with 100 micrograms/ml of oil fly ash decreased lactoferrin binding. Comparable to the particle, exposure of BEAS-2B cells to either 1.0 mM vanadyl sulfate or 1.0 mM iron (III) sulfate, but not to nickel sulfate, for 45 min elevated LfR activity. We conclude that LfRs on respiratory epithelial cells increased after exposure to metal. LfRs could participate in decreasing the oxidative stress presented to the lower respiratory tract by complexing catalytically active metals.
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Bronquios/metabolismo , Metales/farmacología , Receptores de Superficie Celular/efectos de los fármacos , Receptores de Superficie Celular/metabolismo , Contaminación del Aire , Bronquios/citología , Bronquios/efectos de los fármacos , Carbono/farmacología , Línea Celular Transformada , Quelantes/farmacología , Ceniza del Carbón , Medios de Cultivo/química , Medios de Cultivo/farmacología , Deferoxamina/farmacología , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Humanos , Hierro/administración & dosificación , Hierro/farmacología , Lactoferrina/metabolismo , Material Particulado , Inhibidores de la Síntesis de la Proteína/farmacología , Vanadio/farmacologíaRESUMEN
One potential mechanism of injury mediated by air pollution particles is through metal-catalyzed oxidant generation. In one emission source particle, soluble metals have been associated with biological effect and toxicity. However, a majority of metals in ambient air pollution particles can be associated with insoluble components. We tested the hypothesis that concentrations of catalytically active metal in ambient air pollution particles are not equivalent to the concentrations of water-soluble metal. Twelve filters collected from the North Provo, UT, monitoring station were agitated in deionized water. Both the aqueous extract and pellet were isolated, lyophilized, and defined as the water-soluble and insoluble fractions, respectively. The fractions were chemically characterized and ionizable concentrations of metals were measured using inductively coupled plasma emission spectroscopy. While the water-soluble fraction had significantly greater concentrations of ionizable metals per unit mass, the insoluble fraction also had measurable quantities. In vitro oxidant generation by the two fractions, measured as thiobarbituric acid-reactive-products of deoxyribose, corresponded to the concentrations of ionizable rather than total metals. The release of interleukin-8 by cultured respiratory epithelial cells after incubation with the two fractions also coincided with the ionizable metal concentrations. Finally, neutrophil influx and lavage protein levels 24 h after instillation of the two fractions in rats reflected the ionizable metal concentrations, in vitro oxidative stress, and mediator release. We conclude that catalytically active metals can be measured in both the soluble and insoluble fractions of an ambient air pollution particle. These metals corresponded to the biological activity of the two fractions. While in greater concentration in the water-soluble fraction, larger total quantities of catalytically and biologically active metals are likely to be associated with the insoluble fraction as a result of the abundance of the latter.
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Contaminantes Atmosféricos/toxicidad , Metales/toxicidad , Estrés Oxidativo/efectos de los fármacos , Contaminantes Atmosféricos/química , Animales , Bronquios/citología , Bronquios/metabolismo , Líquido del Lavado Bronquioalveolar/química , Líquido del Lavado Bronquioalveolar/citología , Catálisis , Recuento de Células , Células Cultivadas , Humanos , Exposición por Inhalación/efectos adversos , Interleucina-8/metabolismo , Masculino , Metales/química , Neutrófilos/efectos de los fármacos , Oxidantes/química , Ratas , Ratas Sprague-Dawley , Solubilidad , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismoRESUMEN
The metabolism of arachidonic acid can be catalyzed by either one of two enzyme families; the cyclooxygenases or the lipoxygenases. The family of lipoxygenases is divided into four subtypes according to tissue distribution; 5-, 8-, 12-, and 15-lipoxygenase. 12-lipoxygenase metabolites, such as 12(S)-hydroxyeicosatetraenoic acid, have been found to play a central role in the various stages of the metastatic process in tumors and are, therefore, potential targets for anticancer treatment. A variety of lipoxygenase inhibitors already exist and can be classified into five major categories according to their mechanism of inhibition. These include antioxidants, iron chelators, substrate analogues, lipoxygenase-activating protein inhibitors, and, finally, epidermal growth factor-receptor inhibitors.
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Araquidonato 12-Lipooxigenasa/clasificación , Inhibidores Enzimáticos/farmacología , Ácido 12-Hidroxi-5,8,10,14-Eicosatetraenoico/fisiología , Araquidonato 12-Lipooxigenasa/metabolismo , Plaquetas/enzimología , Humanos , Inhibidores de la Lipooxigenasa , Metástasis de la NeoplasiaRESUMEN
We have previously shown that in vitro exposure to metallic compounds enhances expression of interleukin (IL)-6, IL-8, and tumor necrosis factor-alpha in human bronchial epithelial cells. To characterize signaling pathways involved in metal-induced expression of inflammatory mediators and to identify metals that activate them, we studied the effects of As, Cr, Cu, Fe, Ni, V, and Zn on the mitogen-activated protein kinases (MAPK) extracellular receptor kinase (ERK), c-Jun NH2-terminal kinase (JNK), and P38 in BEAS cells. Noncytotoxic concentrations of As, V, and Zn induced a rapid phosphorylation of MAPK in BEAS cells. Activity assays confirmed marked activation of ERK, JNK, and P38 in BEAS cells exposed to As, V, and Zn. Cr and Cu exposure resulted in a relatively small activation of MAPK, whereas Fe and Ni did not activate MAPK under these conditions. Similarly, the transcription factors c-Jun and ATF-2, substrates of JNK and P38, respectively, were markedly phosphorylated in BEAS cells treated with As, Cr, Cu, V, and Zn. The same acute exposure to As, V, or Zn that activated MAPK was sufficient to induce a subsequent increase in IL-8 protein expression in BEAS cells. These data suggest that MAPK may mediate metal-induced expression of inflammatory proteins in human bronchial epithelial cells.